Human amniotic epithelial cells cultured in substitute serum medium maintain their stem cell characteristics for up to four passages

在替代血清培养基中培养的人类羊膜上皮细胞可维持其干细胞特征长达四代

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作者:Ayelet Evron, Shlomit Goldman, Eliezer Shalev

Conclusions

We can conclude that, hAECs maintain their stem cell characteristics when cultured in SSM for up to 4 passages. This makes possible future clinical applications of these cells more feasible.

Results

Purity validation and characterization of freshly isolated and cultured hAECs was assessed through the expression of stem cell specific markers by RT-PCR (gene expression), by immunofluorescence staining and FACS (protein expression). Furthermore, karyotype was performed at passage four in order to exclude possible chromosome anomalies in hAECs cultured in SSM. The differentiation potential of hAECs into the cardiomyogenic lineage was tested through cardiac Troponin T expression by immunohistochemistry. hAECs cultured in SSM maintained expression of all the major pluripotent genes Sox-2, Oct-4 and Nanog as well as the expression of the embryonic stem cell specific surface antigens SSEA-4, SSEA-3 and TRA-1-60 over four passages. Using cardiac differentiation medium containing 10% serum substitute supplement, hAECs differentiated into cardiac troponin T expressing cells. Conclusions: We can conclude that, hAECs maintain their stem cell characteristics when cultured in SSM for up to 4 passages. This makes possible future clinical applications of these cells more feasible.

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